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Differential expression of copper-zinc superoxide dismutase gene of Polygonum sibiricum leaves, stems and underground stems, subjected to high-salt stress.

Qu CP, Xu ZR, Liu GJ, Liu C, Li Y, Wei ZG, Liu GF - Int J Mol Sci (2010)

Bottom Line: Expression analysis by real-time quantitative PCR reveals that the PS-CuZnSOD gene is expressed in leaves, stems and underground stems.PS-CuZnSOD gene expression can be induced by 3% NaHCO(3).The different mRNA levels' expression of PS-CuZnSOD show the gene's different expression modes in leaves, stems and underground stems under the salinity-alkalinity stress.

View Article: PubMed Central - PubMed

Affiliation: The Laboratory of Forest Genetics and Breeding and Biotechnology of Ministry of Education, Northeast Forestry University, 26 Hexing Road, Harbin 150040, China; E-Mails: qcp0451@msn.com.cn (C.-P.Q.); liuchun314@yahoo.com.cn (C.L.); liyangzaixian362@163.com (Y.L.); zhigangwe@163.com (Z.-G.W.); liuguifeng@126.com (G.-F.L.).

ABSTRACT
In aerobic organisms, protection against oxidative damage involves the combined action of highly specialized antioxidant enzymes, such as copper-zinc superoxide dismutase. In this work, a cDNA clone which encodes a copper-zinc superoxide dismutase gene, named PS-CuZnSOD, has been identified from P. sibiricum Laxm. by the rapid amplification of cDNA ends method (RACE). Analysis of the nucleotide sequence reveals that the PS-CuZnSOD gene cDNA clone consists of 669 bp, containing 87 bp in the 5' untranslated region; 459 bp in the open reading frame (ORF) encoding 152 amino acids; and 123 bp in 3' untranslated region. The gene accession nucleotide sequence number in GenBank is GQ472846. Sequence analysis indicates that the protein, like most plant superoxide dismutases (SOD), includes two conserved ecCuZnSOD signatures that are from the amino acids 43 to 51, and from the amino acids 137 to 148, and it has a signal peptide extension in the front of the N-terminus (1-16 aa). Expression analysis by real-time quantitative PCR reveals that the PS-CuZnSOD gene is expressed in leaves, stems and underground stems. PS-CuZnSOD gene expression can be induced by 3% NaHCO(3). The different mRNA levels' expression of PS-CuZnSOD show the gene's different expression modes in leaves, stems and underground stems under the salinity-alkalinity stress.

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The change of PS-CuZnSOD after 3% NaHCO3 exposure in leaves, stems, underground stems’ organs. Total RNA was prepared using SDS reagent for all P. sibiricum Laxm. samples taken at 0, 4, 8, 24, 48, 72, 144 h, independent of 3% NaHCO3 stress condition. After digested with DNase I to eliminate the genome contamination, the cDNA was synthesized using the oligo d (T) primer and random 6 primer. Real-time PCR was performed with the DNA Engine Opticon™-II sequence detection system. SYBR green Real-time PCR mix (TaKaRa) was used for PCR. (A) The expression of PS-CuZnSOD gene in leaves, stems, underground stems’ organ without stress comparison; (B) The levels of PS-CuZnSOD mRNA in leaves tissues; (C) The level of PS-CuZnSOD mRNA in stems tissues; (D) The levels of PS-CuZnSOD mRNA in underground stems tissues. A multiple comparisons test was conducted to compare significant differences in PS-CuZnSOD expression between leaves, stems and underground stems using the SPSS software. A significant level of p = 0.05 was chosen.
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f4-ijms-11-05234: The change of PS-CuZnSOD after 3% NaHCO3 exposure in leaves, stems, underground stems’ organs. Total RNA was prepared using SDS reagent for all P. sibiricum Laxm. samples taken at 0, 4, 8, 24, 48, 72, 144 h, independent of 3% NaHCO3 stress condition. After digested with DNase I to eliminate the genome contamination, the cDNA was synthesized using the oligo d (T) primer and random 6 primer. Real-time PCR was performed with the DNA Engine Opticon™-II sequence detection system. SYBR green Real-time PCR mix (TaKaRa) was used for PCR. (A) The expression of PS-CuZnSOD gene in leaves, stems, underground stems’ organ without stress comparison; (B) The levels of PS-CuZnSOD mRNA in leaves tissues; (C) The level of PS-CuZnSOD mRNA in stems tissues; (D) The levels of PS-CuZnSOD mRNA in underground stems tissues. A multiple comparisons test was conducted to compare significant differences in PS-CuZnSOD expression between leaves, stems and underground stems using the SPSS software. A significant level of p = 0.05 was chosen.

Mentions: Copper-zinc superoxide dismutase expressed in each organ of P. sibiricum Laxm. is shown in Figure 4. In a RT-PCR study, specific primers (SOD-F: 5′-AGTGCGGGAGTTAGTGG-3′ and SOD-R: 5′-CGATGCTCGTCTTCTGG-3′) were used to amplify a 203 bp fragment with cDNA from leaves, stems and underground stems, organs using 18S as a positive control. The RT-PCR showed that the CuZnSOD was detected in leaves, stems and underground stems. In leaves, the increase of the copper-zinc superoxide dismutase mRNA expression level reached its peak in 24 hours after 3% NaHCO3 stress, and gradually decreased (Figure 4B). In stems, the increase of the copper-zinc superoxide dismutase mRNA expression reached its peak in 72 hours after salinity-alkalinity stress (Figure 4C). That is, in leaves and stems they were up-regulated and then down-regulated during 3% NaHCO3 stress. Contrastingly, the copper-zinc superoxide dismutase transcripts were fluctuated and down-regulated after 3% NaHCO3 stress in underground stems’ organs (Figure 4D).


Differential expression of copper-zinc superoxide dismutase gene of Polygonum sibiricum leaves, stems and underground stems, subjected to high-salt stress.

Qu CP, Xu ZR, Liu GJ, Liu C, Li Y, Wei ZG, Liu GF - Int J Mol Sci (2010)

The change of PS-CuZnSOD after 3% NaHCO3 exposure in leaves, stems, underground stems’ organs. Total RNA was prepared using SDS reagent for all P. sibiricum Laxm. samples taken at 0, 4, 8, 24, 48, 72, 144 h, independent of 3% NaHCO3 stress condition. After digested with DNase I to eliminate the genome contamination, the cDNA was synthesized using the oligo d (T) primer and random 6 primer. Real-time PCR was performed with the DNA Engine Opticon™-II sequence detection system. SYBR green Real-time PCR mix (TaKaRa) was used for PCR. (A) The expression of PS-CuZnSOD gene in leaves, stems, underground stems’ organ without stress comparison; (B) The levels of PS-CuZnSOD mRNA in leaves tissues; (C) The level of PS-CuZnSOD mRNA in stems tissues; (D) The levels of PS-CuZnSOD mRNA in underground stems tissues. A multiple comparisons test was conducted to compare significant differences in PS-CuZnSOD expression between leaves, stems and underground stems using the SPSS software. A significant level of p = 0.05 was chosen.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC3100833&req=5

f4-ijms-11-05234: The change of PS-CuZnSOD after 3% NaHCO3 exposure in leaves, stems, underground stems’ organs. Total RNA was prepared using SDS reagent for all P. sibiricum Laxm. samples taken at 0, 4, 8, 24, 48, 72, 144 h, independent of 3% NaHCO3 stress condition. After digested with DNase I to eliminate the genome contamination, the cDNA was synthesized using the oligo d (T) primer and random 6 primer. Real-time PCR was performed with the DNA Engine Opticon™-II sequence detection system. SYBR green Real-time PCR mix (TaKaRa) was used for PCR. (A) The expression of PS-CuZnSOD gene in leaves, stems, underground stems’ organ without stress comparison; (B) The levels of PS-CuZnSOD mRNA in leaves tissues; (C) The level of PS-CuZnSOD mRNA in stems tissues; (D) The levels of PS-CuZnSOD mRNA in underground stems tissues. A multiple comparisons test was conducted to compare significant differences in PS-CuZnSOD expression between leaves, stems and underground stems using the SPSS software. A significant level of p = 0.05 was chosen.
Mentions: Copper-zinc superoxide dismutase expressed in each organ of P. sibiricum Laxm. is shown in Figure 4. In a RT-PCR study, specific primers (SOD-F: 5′-AGTGCGGGAGTTAGTGG-3′ and SOD-R: 5′-CGATGCTCGTCTTCTGG-3′) were used to amplify a 203 bp fragment with cDNA from leaves, stems and underground stems, organs using 18S as a positive control. The RT-PCR showed that the CuZnSOD was detected in leaves, stems and underground stems. In leaves, the increase of the copper-zinc superoxide dismutase mRNA expression level reached its peak in 24 hours after 3% NaHCO3 stress, and gradually decreased (Figure 4B). In stems, the increase of the copper-zinc superoxide dismutase mRNA expression reached its peak in 72 hours after salinity-alkalinity stress (Figure 4C). That is, in leaves and stems they were up-regulated and then down-regulated during 3% NaHCO3 stress. Contrastingly, the copper-zinc superoxide dismutase transcripts were fluctuated and down-regulated after 3% NaHCO3 stress in underground stems’ organs (Figure 4D).

Bottom Line: Expression analysis by real-time quantitative PCR reveals that the PS-CuZnSOD gene is expressed in leaves, stems and underground stems.PS-CuZnSOD gene expression can be induced by 3% NaHCO(3).The different mRNA levels' expression of PS-CuZnSOD show the gene's different expression modes in leaves, stems and underground stems under the salinity-alkalinity stress.

View Article: PubMed Central - PubMed

Affiliation: The Laboratory of Forest Genetics and Breeding and Biotechnology of Ministry of Education, Northeast Forestry University, 26 Hexing Road, Harbin 150040, China; E-Mails: qcp0451@msn.com.cn (C.-P.Q.); liuchun314@yahoo.com.cn (C.L.); liyangzaixian362@163.com (Y.L.); zhigangwe@163.com (Z.-G.W.); liuguifeng@126.com (G.-F.L.).

ABSTRACT
In aerobic organisms, protection against oxidative damage involves the combined action of highly specialized antioxidant enzymes, such as copper-zinc superoxide dismutase. In this work, a cDNA clone which encodes a copper-zinc superoxide dismutase gene, named PS-CuZnSOD, has been identified from P. sibiricum Laxm. by the rapid amplification of cDNA ends method (RACE). Analysis of the nucleotide sequence reveals that the PS-CuZnSOD gene cDNA clone consists of 669 bp, containing 87 bp in the 5' untranslated region; 459 bp in the open reading frame (ORF) encoding 152 amino acids; and 123 bp in 3' untranslated region. The gene accession nucleotide sequence number in GenBank is GQ472846. Sequence analysis indicates that the protein, like most plant superoxide dismutases (SOD), includes two conserved ecCuZnSOD signatures that are from the amino acids 43 to 51, and from the amino acids 137 to 148, and it has a signal peptide extension in the front of the N-terminus (1-16 aa). Expression analysis by real-time quantitative PCR reveals that the PS-CuZnSOD gene is expressed in leaves, stems and underground stems. PS-CuZnSOD gene expression can be induced by 3% NaHCO(3). The different mRNA levels' expression of PS-CuZnSOD show the gene's different expression modes in leaves, stems and underground stems under the salinity-alkalinity stress.

Show MeSH
Related in: MedlinePlus